Important role of mandibular organ in molting, growth, and survival of mud crabscylla olivacea

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1 International Journal of ChemTech Research CODEN (USA): IJCRGG, ISSN: , ISSN(Online): Vol.9 No.12, pp , 2016 Important role of mandibular organ in molting, growth, and survival of mud crabscylla olivacea Akbar Marzuki Tahya 1, Muhammad Zairin Jr 1 *, Arief Boediono 2 I Made Artika 3, and Muhammad Agus Suprayudi 1. 1 Department of Aquaculture, Bogor Agricultural University, Indonesia 2 Department of Anatomy, Physiology, Pharmacology, Bogor Agricultural University, Indonesia 3 Department of Biochemistry, Bogor Agricultural University, Indonesia Abstract : Molting is a long process in crustaceans, it is substantially associated to the growth, reproduction, and survival. The physiological of molting is essentially linked to the hormonal control. Henceforth a clear knowledge about molting process and hormonal control are important. Mandibular organ (MO) is one of many organs thats play role in molting and reproduction. The influence of exogenous MO in the progress of molt, growth, and mortality in commercial mud crab was investigated. Injection of MO into intermolt crabs increased molting percentages, molting simultaneity, growth acceleration, and adaptability. The results confirm that the MO is involved in the control of molting in Scylla olivacea. Nonetheless, further research is required to study the role of exogenous MO in the adaptability of crabs. Keywords.Adaptability; Crab; Growth; Injection; Mandibular Organ; Molting. Introduction The regulation of molting in insects and crustacean can be accelerated by multy-factors. The presence of endogenous hormone is one of the internal factors, which can initiate premolt. Premolt is one of molting phases. This phase is crucial caused of conciseness in energy preparation to face ecdysis. Molting is physiological-complexity that required many mechanisms. Mandibular Organ (MO) is one of many organs thats play role in molting (1;2). MO firstly described as another pair of crustacean glands by LeRoux (3). Formerly, MO and Y-organ were hard to discern because the similarity both in histology and anatomy. Since MO is known as Methyl Farnesoate (MF) producing-organ, now biochemically can be distinguish from Y-organ (4). In aquaculture, molting acceleration and simultaneous are required because most of the reared crabs are incapable to do that. In the present study, we investigated the role of MO in molting progress of commercial crab S. olivacea. Material and Method Male crabs Scylla olivacea were obtained from fisherman in South Sulawesi Province, Indonesia. Healthy crabs and ± 100 gram of body weight (BW) were selected as test crabs. The crabs were maintained individual in crab box and fed fresh fish 10% of crab s body weight per day. The crabs were divided into 2 groups, control (A) and injected of MO (I).

2 Muhammad Zairin Jr et al /International Journal of ChemTech Research, 2016,9(12): Result and Discussion Molting The injection of mandibular organ increased the molting percentage and was found to have an effect on simultaneity of molting. The present study showed that the injected crabs had higher percentage of molting than control test crabs (Figure 1). Figure 1. Molting percentages of control (A) and injected crabs (I) Figure 2. The time required for molting of test crabs

3 Muhammad Zairin Jr et al /International Journal of ChemTech Research, 2016,9(12): The results showed that the injected of MO had higher molting percentage i.e. 80, 80, and 90%, than control crabs i.e. 20, 10, and 50%. The results presented in Figure 1 also show that the percentage values of injected crabs are more consistent than control crabs. The role of MO has been observed in several crustaceans, such as Oziotelphusa senex senex (4;5), Procambarus clarkii (6), Charybdis lucifera (7). The results confirm that the molting is under the positive regulation of MO. Therefore, suggested that MF of MO play an important role for stimulation of Y-organ. When stimulation through MO extraction are administrated, the ecdysteroid level increases and releases in haemolymph stream that may be responsible for simultaneous molting (Figure 2). In control crabs the time required for molting between 20 to 41 days to reach of 26.67%, while in injected required shorter time i.e 26 to 34 days to reach the percentage of 83.33%. In soft shell crab farming, rapidity and simultaneity of molting is expected in order that the production process can be achieved early and efficient of maintenance. The present study showed that control crabs were faster molting then injected. While the injected of MO more simultaneous of molting. The longer time in the control treatment was reasonable due unavailability trigger of ecdysteroid to be secreted out from the Y organ. Unlike the injection treatment of MO which has MF as a trigger. As MF administration resulting response of molting in Cherax quadricarinatus (8), male and female of O. senex senex (5). Injection of MO was effectifely stimulated ecdysteroid which resulted in simultaneity of molting. If it follows the normal rules without stimulation, the release of MF from MO will be affected by control of X Organ Sinus Gland (XOSG). The Mandibular Organ Inhibiting Hormone (MOIH) from XOSG will be influence to molting of crab, because the role of MOIH is negatively control MO to secrete and release of MF to hemolymph, as the result that molting will be postpone. Molting process also will be delay, when Molting Inhibiting Hormones (MIH)s from XOSG are widely circulated in hemolymph. Growth In Figure 3, data on the body weight (BW) and the carapace width (CW) of crabs are reported. As shown, the CW increased with increasing BW in both postmolt growth and non-molt growth. Based on this study, the BW increased on an average by g and CW increased on an average by mm in postmolt growth of control, and BW increased on an average by g and CW increased on average by mm in postmolt growth of injected. While in non-molt growth, the BW increased on an average by 7.77 g and CW increased on an average 3.91 mm in control, and BW increased on an average by 9.25 g and CW increased on an average 3.98 mm in injected. Figure 3. Success molting (postmolt) and non-molt growth of testing crabs. During 45 days observation, BW and CW increased both in control and injection. But BW and CW of crabs increased significantly in postmolt growth. The administration of MO may stimulates the source of molting hormone, resulting in increased ecdysteroid level and internal body growth. In fresh water crab O. Senex senex, the administration of MF has been reported to stimulate testis growth (5). Weight difference that occurs can not be separated from the role of MF and ecdysteroid, such as in protein synthesis (9), and molting (4;8). However, at the end of the observation we found some test crabs were unsuccessfully molting, but the data of BW and CW were taken. Based on data, it needs to be pointed out that the growth also occured although in nonmolting crabs, but growth of non-molt crabs increased with a low percentage. The significant growth in postmolt and lack of growth in non-molt crabs are affected by multy factors. Kleinholz (10) reviewed that molting in crustaceans is affected by extrinsic factors i.e. temperature, salinity of medium, light intensity,

4 Muhammad Zairin Jr et al /International Journal of ChemTech Research, 2016,9(12): pollutants, and by intrinsic factors i.e. nutritional, and hormones. Based on swim legs observation, the control crabs groups were also in intermolt stage after the completion of experiment (Figure 4.A). All of the non-molt crabs, which received the MO injection entered in to premolt stage (Figure 4.I). These observation ilustrate the role of MO on molting progress in mud crabs S. olivacea. Much evidence shows that MF in MO play a positive role in the control of molting and growth, but relatively few data have been provided relating the physiological role of MF. Figure 4. Swim legs observation of control (A) and injection (I), non-molt crabs, the arrow indicated retraction Mortality The mortality during the observation occured in control crabs. Figure 5, indicates that the mortalities of crabs only occured in control, and all of injected survived for 45 days observation. The control crabs in A1, A2, and A3 died by 30, 20, and 20%, respectively. It is interesting to note that mortality consistently occured in the control crabs. This may be caused by the stresses of cultivation environment, and then mortality started on day 2 to 31. The present study showed that the injected crabs more adaptable to environmental stress and acclimation. It needs to be pointed out that molting failure syndrome can be also be occured by the fit and proper of health. So it can be assumed that the test crabs were in a healthy and ready for molting. Lovett et al. (11; 12; 13) found an increase of MF in Carcinus maenas exposed to environmental stress. Further research is needed to study the MF of MO in the adaptability of crabs. Figure 5. Mortality percentages during observation.

5 Muhammad Zairin Jr et al /International Journal of ChemTech Research, 2016,9(12): Conclusion The results of this study indicated that the role of MO in molting progress, molting simultaneity, growth acceleration, and adaptability of commercial mud crabs Scylla olivacea. These results provide strong evidence that MF play an important role in physiology of molting. Acknowledgements This study was supported by Government of the Republic of Indonesia. The authors are gratefull to Professor Yushinta Fujaya, and Faculty of Marine Science and Fishery of Hasanuddin University Makassar for facilitating in Crab s Pond. References 1. Tahya AM, Zairin MJr, Boediono A, Artika IM, Suprayudi MA Expression of RNA encode FAMeT in mandibular organ of mud crabs Scylla olivacea. IJ Pharmtech Resear. 9(3): Sunarti Y, Soejoedono RD, Mayasari NLPI, Tahya AM RNA expression of farnesoic acid O- methyltransferase in mandibular organ of intermolt and premolt mud crabs Scylla olivacea. AACL Bioflux. 9(2): Le Roux A Description d organesmandibulaires nouveaux chez les crustacedecapodes. C. R. Academy of Science Paris. 266, Nagaraju GPC, Reddy PR, Reddy PS Mandibular organ: it s relation to body weight, sex, molt and reproduction in the crab,oziotelphusasenexsenexfabricus (1791). Aquaculture 232, Reddy PR, Nagaraju GPC, Reddy PS Involvement of methyl farnesoate in the regulation of molting and reproduction in the freshwater crab Oziotelphusasenexsenex. Journal of Crustacean Biology 24, Laufer H, Demir N, Pan X, Stuart JD, Ahl JSB Methyl farnesoate controls adult male morphogenesis in the crayfish, Procambarusclarkii. J. Insect Physiol. 51, Allayie SA, Ravichandran S, Bhat BA Role of mandibular glands in growth of mangrove crab, Charybdis lucifera (Fabricius, 1798). World Journal of Zoology 5(2), Abdu U, Barki A, Karplus I, Barel S, Takac P, Yehezkel G, Laufer H, Sagi A Physiological effect of methyl farnesoate and pyriproxyfen on wintering female crayfish Cheraxquadricarinatus. Aquaculture 202, Paulson CR, Skinner DM Molecular action of 20-hydroxyecdysone, methyl farnesoate, and juvenile hormone on crab tissues. American zoologist 28, 83A. 10. Kleinholz LH Biochemistry of crustacean hormones. In Bliss, D.E., Mantel, L.H. (Eds.), TheBiology of Crustacea, Integument Pigments and Hormonal Process, vol. 9. Academic Press, NewYork, pp Lovett DL, Clifford PD, Borst DW Physiological stresselevateshemolymph levels of methyl farnesoate in the green crabcarcinusmaenas. Biol. Bull. 193, Lovett DL, Verzi MP, Clifford PD, Borst DW Hemolymph levels of methyl farnesoate increase in response toosmotic stress in the green crab Carcinusmaenas. Comp.Biochem. Physiol. 128, Lovett DL, Tanner CA, Glomski K, Ricart TM, Borst DW The effect of seawater composition and osmolality onhemolymph levels of methyl farnesoate in the green crab Carcinusmaenas. Comp. Biochem. Physiol. 143, *****